One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV
Identifieur interne : 000C78 ( Pmc/Curation ); précédent : 000C77; suivant : 000C79One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV
Auteurs : Se Hee Lee [Corée du Sud] ; Yun Hee Baek [Corée du Sud] ; Yang-Hoon Kim [Corée du Sud] ; Young-Ki Choi [Corée du Sud] ; Min-Suk Song [Corée du Sud] ; Ji-Young Ahn [Corée du Sud]Source :
- Frontiers in Microbiology [ 1664-302X ] ; 2017.
Abstract
Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U
Url:
DOI: 10.3389/fmicb.2016.02166
PubMed: 28119682
PubMed Central: 5220095
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000C78
Links to Exploration step
PMC:5220095Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV</title><author><name sortKey="Lee, Se Hee" sort="Lee, Se Hee" uniqKey="Lee S" first="Se Hee" last="Lee">Se Hee Lee</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Baek, Yun Hee" sort="Baek, Yun Hee" uniqKey="Baek Y" first="Yun Hee" last="Baek">Yun Hee Baek</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Kim, Yang Hoon" sort="Kim, Yang Hoon" uniqKey="Kim Y" first="Yang-Hoon" last="Kim">Yang-Hoon Kim</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Choi, Young Ki" sort="Choi, Young Ki" uniqKey="Choi Y" first="Young-Ki" last="Choi">Young-Ki Choi</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Song, Min Suk" sort="Song, Min Suk" uniqKey="Song M" first="Min-Suk" last="Song">Min-Suk Song</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Ahn, Ji Young" sort="Ahn, Ji Young" uniqKey="Ahn J" first="Ji-Young" last="Ahn">Ji-Young Ahn</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">28119682</idno><idno type="pmc">5220095</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5220095</idno><idno type="RBID">PMC:5220095</idno><idno type="doi">10.3389/fmicb.2016.02166</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">000C78</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000C78</idno><idno type="wicri:Area/Pmc/Curation">000C78</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000C78</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV</title><author><name sortKey="Lee, Se Hee" sort="Lee, Se Hee" uniqKey="Lee S" first="Se Hee" last="Lee">Se Hee Lee</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Baek, Yun Hee" sort="Baek, Yun Hee" uniqKey="Baek Y" first="Yun Hee" last="Baek">Yun Hee Baek</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Kim, Yang Hoon" sort="Kim, Yang Hoon" uniqKey="Kim Y" first="Yang-Hoon" last="Kim">Yang-Hoon Kim</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Choi, Young Ki" sort="Choi, Young Ki" uniqKey="Choi Y" first="Young-Ki" last="Choi">Young-Ki Choi</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Song, Min Suk" sort="Song, Min Suk" uniqKey="Song M" first="Min-Suk" last="Song">Min-Suk Song</name><affiliation wicri:level="1"><nlm:aff id="aff2"><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Ahn, Ji Young" sort="Ahn, Ji Young" uniqKey="Ahn J" first="Ji-Young" last="Ahn">Ji-Young Ahn</name><affiliation wicri:level="1"><nlm:aff id="aff1"><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></nlm:aff><country xml:lang="fr">Corée du Sud</country><wicri:regionArea></wicri:regionArea></affiliation></author></analytic><series><title level="j">Frontiers in Microbiology</title><idno type="eISSN">1664-302X</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U <italic>Bst</italic> polymerase, implying that the reaction was able to detect four infectious viral genome copies of MERS-CoV within a 60 min reaction time period. Significantly, EvaGreen dye has better signal read-out properties in one-pot RT-LAMP reaction and is more compatible with DNA polymerase than SYBR green I. Isothermally amplified specific N genes were further evaluated using field-deployable microchamber devices, leading to the specific identification of as few as 0.4 infectious viral genome copies, with no cross-reaction to the other acute respiratory disease viruses, including influenza type A (H1N1 and H3N2), type B, human coronavirus 229E, and human metapneumovirus. This sensitive, specific and feasible method provides a large-scale technical support in emergencies, and is also applied as a sample-to-detection module in Point of Care Testing devices.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Abd El Wahed, A" uniqKey="Abd El Wahed A">A. Abd El Wahed</name></author><author><name sortKey="Patel, P" uniqKey="Patel P">P. Patel</name></author><author><name sortKey="Heidenreich, D" uniqKey="Heidenreich D">D. Heidenreich</name></author><author><name sortKey="Hufert, F T" uniqKey="Hufert F">F. T. Hufert</name></author><author><name sortKey="Weidmann, M" uniqKey="Weidmann M">M. Weidmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alagaili, A N" uniqKey="Alagaili A">A. N. Alagaili</name></author><author><name sortKey="Briese, T" uniqKey="Briese T">T. Briese</name></author><author><name sortKey="Mishra, N" uniqKey="Mishra N">N. Mishra</name></author><author><name sortKey="Kapoor, V" uniqKey="Kapoor V">V. Kapoor</name></author><author><name sortKey="Sameroff, S C" uniqKey="Sameroff S">S. C. Sameroff</name></author><author><name sortKey="Burbelo, P D" uniqKey="Burbelo P">P. D. Burbelo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Aoi, Y" uniqKey="Aoi Y">Y. Aoi</name></author><author><name sortKey="Hosogai, M" uniqKey="Hosogai M">M. Hosogai</name></author><author><name sortKey="Tsuneda, S" uniqKey="Tsuneda S">S. Tsuneda</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arezi, B" uniqKey="Arezi B">B. Arezi</name></author><author><name sortKey="Hogrefe, H" uniqKey="Hogrefe H">H. Hogrefe</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arunrut, N" uniqKey="Arunrut N">N. Arunrut</name></author><author><name sortKey="Kampeera, J" uniqKey="Kampeera J">J. Kampeera</name></author><author><name sortKey="Sirithammajak, S" uniqKey="Sirithammajak S">S. Sirithammajak</name></author><author><name sortKey="Sanguanrut, P" uniqKey="Sanguanrut P">P. Sanguanrut</name></author><author><name sortKey="Proespraiwong, P" uniqKey="Proespraiwong P">P. Proespraiwong</name></author><author><name sortKey="Suebsing, R" uniqKey="Suebsing R">R. Suebsing</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bhadra, S" uniqKey="Bhadra S">S. Bhadra</name></author><author><name sortKey="Jiang, Y S" uniqKey="Jiang Y">Y. S. Jiang</name></author><author><name sortKey="Kumar, M R" uniqKey="Kumar M">M. R. Kumar</name></author><author><name sortKey="Johnson, R F" uniqKey="Johnson R">R. F. Johnson</name></author><author><name sortKey="Hensley, L E" uniqKey="Hensley L">L. E. Hensley</name></author><author><name sortKey="Ellington, A D" uniqKey="Ellington A">A. D. Ellington</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bialek, S R" uniqKey="Bialek S">S. R. Bialek</name></author><author><name sortKey="Allen, D" uniqKey="Allen D">D. Allen</name></author><author><name sortKey="Alvarado Ramy, F" uniqKey="Alvarado Ramy F">F. Alvarado-Ramy</name></author><author><name sortKey="Arthur, R" uniqKey="Arthur R">R. Arthur</name></author><author><name sortKey="Balajee, A" uniqKey="Balajee A">A. Balajee</name></author><author><name sortKey="Bell, D" uniqKey="Bell D">D. Bell</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bin, S Y" uniqKey="Bin S">S. Y. Bin</name></author><author><name sortKey="Heo, J Y" uniqKey="Heo J">J. Y. Heo</name></author><author><name sortKey="Song, M S" uniqKey="Song M">M. S. Song</name></author><author><name sortKey="Lee, J" uniqKey="Lee J">J. Lee</name></author><author><name sortKey="Kim, E H" uniqKey="Kim E">E. H. Kim</name></author><author><name sortKey="Park, S J" uniqKey="Park S">S. J. Park</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cai, T" uniqKey="Cai T">T. Cai</name></author><author><name sortKey="Lou, G" uniqKey="Lou G">G. Lou</name></author><author><name sortKey="Yang, J" uniqKey="Yang J">J. Yang</name></author><author><name sortKey="Xu, D" uniqKey="Xu D">D. Xu</name></author><author><name sortKey="Meng, Z" uniqKey="Meng Z">Z. Meng</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Corman, V M" uniqKey="Corman V">V. M. Corman</name></author><author><name sortKey="Eckerle, I" uniqKey="Eckerle I">I. Eckerle</name></author><author><name sortKey="Bleicker, T" uniqKey="Bleicker T">T. Bleicker</name></author><author><name sortKey="Zaki, A" uniqKey="Zaki A">A. Zaki</name></author><author><name sortKey="Landt, O" uniqKey="Landt O">O. Landt</name></author><author><name sortKey="Eschbach Bludau, M" uniqKey="Eschbach Bludau M">M. Eschbach-Bludau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Corman, V M" uniqKey="Corman V">V. M. Corman</name></author><author><name sortKey="Olschlager, S" uniqKey="Olschlager S">S. Olschlager</name></author><author><name sortKey="Wendtner, C M" uniqKey="Wendtner C">C. M. Wendtner</name></author><author><name sortKey="Drexler, J F" uniqKey="Drexler J">J. F. Drexler</name></author><author><name sortKey="Hess, M" uniqKey="Hess M">M. Hess</name></author><author><name sortKey="Drosten, C" uniqKey="Drosten C">C. Drosten</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hong, T C" uniqKey="Hong T">T. C. Hong</name></author><author><name sortKey="Mai, Q L" uniqKey="Mai Q">Q. L. Mai</name></author><author><name sortKey="Cuong, D V" uniqKey="Cuong D">D. V. Cuong</name></author><author><name sortKey="Parida, M" uniqKey="Parida M">M. Parida</name></author><author><name sortKey="Minekawa, H" uniqKey="Minekawa H">H. Minekawa</name></author><author><name sortKey="Notomi, T" uniqKey="Notomi T">T. Notomi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Huang, J" uniqKey="Huang J">J. Huang</name></author><author><name sortKey="Sun, Y" uniqKey="Sun Y">Y. Sun</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Xiao, H" uniqKey="Xiao H">H. Xiao</name></author><author><name sortKey="Zhuang, S" uniqKey="Zhuang S">S. Zhuang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Iwata, S" uniqKey="Iwata S">S. Iwata</name></author><author><name sortKey="Shibata, Y" uniqKey="Shibata Y">Y. Shibata</name></author><author><name sortKey="Kawada, J" uniqKey="Kawada J">J. Kawada</name></author><author><name sortKey="Hara, S" uniqKey="Hara S">S. Hara</name></author><author><name sortKey="Nishiyama, Y" uniqKey="Nishiyama Y">Y. Nishiyama</name></author><author><name sortKey="Morishima, T" uniqKey="Morishima T">T. Morishima</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kaneko, H" uniqKey="Kaneko H">H. Kaneko</name></author><author><name sortKey="Iida, T" uniqKey="Iida T">T. Iida</name></author><author><name sortKey="Aoki, K" uniqKey="Aoki K">K. Aoki</name></author><author><name sortKey="Ohno, S" uniqKey="Ohno S">S. Ohno</name></author><author><name sortKey="Suzutani, T" uniqKey="Suzutani T">T. Suzutani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kuboki, N" uniqKey="Kuboki N">N. Kuboki</name></author><author><name sortKey="Inoue, N" uniqKey="Inoue N">N. Inoue</name></author><author><name sortKey="Sakurai, T" uniqKey="Sakurai T">T. Sakurai</name></author><author><name sortKey="Di Cello, F" uniqKey="Di Cello F">F. Di Cello</name></author><author><name sortKey="Grab, D J" uniqKey="Grab D">D. J. Grab</name></author><author><name sortKey="Suzuki, H" uniqKey="Suzuki H">H. Suzuki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lau, S K" uniqKey="Lau S">S. K. Lau</name></author><author><name sortKey="Li, K S" uniqKey="Li K">K. S. Li</name></author><author><name sortKey="Huang, Y" uniqKey="Huang Y">Y. Huang</name></author><author><name sortKey="Shek, C T" uniqKey="Shek C">C. T. Shek</name></author><author><name sortKey="Tse, H" uniqKey="Tse H">H. Tse</name></author><author><name sortKey="Wang, M" uniqKey="Wang M">M. Wang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, W" uniqKey="Li W">W. Li</name></author><author><name sortKey="Shi, Z" uniqKey="Shi Z">Z. Shi</name></author><author><name sortKey="Yu, M" uniqKey="Yu M">M. Yu</name></author><author><name sortKey="Ren, W" uniqKey="Ren W">W. Ren</name></author><author><name sortKey="Smith, C" uniqKey="Smith C">C. Smith</name></author><author><name sortKey="Epstein, J H" uniqKey="Epstein J">J. H. Epstein</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mailles, A" uniqKey="Mailles A">A. Mailles</name></author><author><name sortKey="Blanckaert, K" uniqKey="Blanckaert K">K. Blanckaert</name></author><author><name sortKey="Chaud, P" uniqKey="Chaud P">P. Chaud</name></author><author><name sortKey="Van Der Werf, S" uniqKey="Van Der Werf S">S. van der Werf</name></author><author><name sortKey="Lina, B" uniqKey="Lina B">B. Lina</name></author><author><name sortKey="Caro, V" uniqKey="Caro V">V. Caro</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mao, F" uniqKey="Mao F">F. Mao</name></author><author><name sortKey="Leung, W Y" uniqKey="Leung W">W. Y. Leung</name></author><author><name sortKey="Xin, X" uniqKey="Xin X">X. Xin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Memish, Z A" uniqKey="Memish Z">Z. A. Memish</name></author><author><name sortKey="Assiri, A M" uniqKey="Assiri A">A. M. Assiri</name></author><author><name sortKey="Al Tawfiq, J A" uniqKey="Al Tawfiq J">J. A. Al-Tawfiq</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mohd, H A" uniqKey="Mohd H">H. A. Mohd</name></author><author><name sortKey="Al Tawfiq, J A" uniqKey="Al Tawfiq J">J. A. Al-Tawfiq</name></author><author><name sortKey="Memish, Z A" uniqKey="Memish Z">Z. A. Memish</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Parida, M" uniqKey="Parida M">M. Parida</name></author><author><name sortKey="Horioke, K" uniqKey="Horioke K">K. Horioke</name></author><author><name sortKey="Ishida, H" uniqKey="Ishida H">H. Ishida</name></author><author><name sortKey="Dash, P K" uniqKey="Dash P">P. K. Dash</name></author><author><name sortKey="Saxena, P" uniqKey="Saxena P">P. Saxena</name></author><author><name sortKey="Jana, A M" uniqKey="Jana A">A. M. Jana</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Parida, M" uniqKey="Parida M">M. Parida</name></author><author><name sortKey="Sannarangaiah, S" uniqKey="Sannarangaiah S">S. Sannarangaiah</name></author><author><name sortKey="Dash, P K" uniqKey="Dash P">P. K. Dash</name></author><author><name sortKey="Rao, P V" uniqKey="Rao P">P. V. Rao</name></author><author><name sortKey="Morita, K" uniqKey="Morita K">K. Morita</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Poon, L L" uniqKey="Poon L">L. L. Poon</name></author><author><name sortKey="Leung, C S" uniqKey="Leung C">C. S. Leung</name></author><author><name sortKey="Chan, K H" uniqKey="Chan K">K. H. Chan</name></author><author><name sortKey="Lee, J H" uniqKey="Lee J">J. H. Lee</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K. Y. Yuen</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y. Guan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shirato, K" uniqKey="Shirato K">K. Shirato</name></author><author><name sortKey="Yano, T" uniqKey="Yano T">T. Yano</name></author><author><name sortKey="Senba, S" uniqKey="Senba S">S. Senba</name></author><author><name sortKey="Akachi, S" uniqKey="Akachi S">S. Akachi</name></author><author><name sortKey="Kobayashi, T" uniqKey="Kobayashi T">T. Kobayashi</name></author><author><name sortKey="Nishinaka, T" uniqKey="Nishinaka T">T. Nishinaka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S. van Boheemen</name></author><author><name sortKey="De Graaf, M" uniqKey="De Graaf M">M. de Graaf</name></author><author><name sortKey="Lauber, C" uniqKey="Lauber C">C. Lauber</name></author><author><name sortKey="Bestebroer, T M" uniqKey="Bestebroer T">T. M. Bestebroer</name></author><author><name sortKey="Raj, V S" uniqKey="Raj V">V. S. Raj</name></author><author><name sortKey="Zaki, A M" uniqKey="Zaki A">A. M. Zaki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, J" uniqKey="Wang J">J. Wang</name></author><author><name sortKey="Chen, Z" uniqKey="Chen Z">Z. Chen</name></author><author><name sortKey="Corstjens, P L" uniqKey="Corstjens P">P. L. Corstjens</name></author><author><name sortKey="Mauk, M G" uniqKey="Mauk M">M. G. Mauk</name></author><author><name sortKey="Bau, H H" uniqKey="Bau H">H. H. Bau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yang, J" uniqKey="Yang J">J. Yang</name></author><author><name sortKey="Yang, R" uniqKey="Yang R">R. Yang</name></author><author><name sortKey="Cheng, A" uniqKey="Cheng A">A. Cheng</name></author><author><name sortKey="Wang, M" uniqKey="Wang M">M. Wang</name></author><author><name sortKey="Fu, L" uniqKey="Fu L">L. Fu</name></author><author><name sortKey="Yang, S" uniqKey="Yang S">S. Yang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zaki, A M" uniqKey="Zaki A">A. M. Zaki</name></author><author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S. van Boheemen</name></author><author><name sortKey="Bestebroer, T M" uniqKey="Bestebroer T">T. M. Bestebroer</name></author><author><name sortKey="Osterhaus, A D" uniqKey="Osterhaus A">A. D. Osterhaus</name></author><author><name sortKey="Fouchier, R A" uniqKey="Fouchier R">R. A. Fouchier</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhang, C" uniqKey="Zhang C">C. Zhang</name></author><author><name sortKey="Xing, D" uniqKey="Xing D">D. Xing</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Front Microbiol</journal-id><journal-id journal-id-type="iso-abbrev">Front Microbiol</journal-id><journal-id journal-id-type="publisher-id">Front. Microbiol.</journal-id><journal-title-group><journal-title>Frontiers in Microbiology</journal-title></journal-title-group><issn pub-type="epub">1664-302X</issn><publisher><publisher-name>Frontiers Media S.A.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28119682</article-id><article-id pub-id-type="pmc">5220095</article-id><article-id pub-id-type="doi">10.3389/fmicb.2016.02166</article-id><article-categories><subj-group subj-group-type="heading"><subject>Microbiology</subject><subj-group><subject>Original Research</subject></subj-group></subj-group></article-categories><title-group><article-title>One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Lee</surname><given-names>Se Hee</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="author-notes" rid="fn002"><sup>†</sup></xref></contrib><contrib contrib-type="author"><name><surname>Baek</surname><given-names>Yun Hee</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref><xref ref-type="author-notes" rid="fn002"><sup>†</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Yang-Hoon</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Choi</surname><given-names>Young-Ki</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Song</surname><given-names>Min-Suk</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref><xref ref-type="author-notes" rid="fn001"><sup>*</sup></xref><uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/384564/overview"></uri></contrib><contrib contrib-type="author"><name><surname>Ahn</surname><given-names>Ji-Young</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="author-notes" rid="fn001"><sup>*</sup></xref><uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/384198/overview"></uri></contrib></contrib-group><aff id="aff1"><sup>1</sup><institution>School of Biological Sciences, Chungbuk National University</institution><country>Cheongju, South Korea</country></aff><aff id="aff2"><sup>2</sup><institution>College of Medicine and Medical Research Institute, Chungbuk National University</institution><country>Cheongju, South Korea</country></aff><author-notes><fn fn-type="edited-by"><p>Edited by: <italic>Masayuki Saijo, National Institute of Infectious Diseases, Japan</italic></p></fn><fn fn-type="edited-by"><p>Reviewed by: <italic>Shuetsu Fukushi, National Institute of Infectious Diseases, Japan; Katsumi Mizuta, Yamagata Prefectural Institute of Public Health, Japan</italic></p></fn><corresp id="fn001">*Correspondence: <italic>Ji-Young Ahn, <email xlink:type="simple">jyahn@chungbuk.ac.kr</email> Min-Suk Song, <email xlink:type="simple">songminsuk@chungbuk.ac.kr</email></italic></corresp><fn fn-type="other" id="fn002"><p><sup>†</sup><italic>These authors have contributed equally to this work.</italic></p></fn><fn fn-type="other" id="fn003"><p>This article was submitted to Virology, a section of the journal Frontiers in Microbiology</p></fn></author-notes><pub-date pub-type="epub"><day>09</day><month>1</month><year>2017</year></pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><volume>7</volume><elocation-id>2166</elocation-id><history><date date-type="received"><day>12</day><month>10</month><year>2016</year></date><date date-type="accepted"><day>23</day><month>12</month><year>2016</year></date></history><permissions><copyright-statement>Copyright © 2017 Lee, Baek, Kim, Choi, Song and Ahn.</copyright-statement><copyright-year>2017</copyright-year><copyright-holder>Lee, Baek, Kim, Choi, Song and Ahn</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p></license></permissions><abstract><p>Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U <italic>Bst</italic> polymerase, implying that the reaction was able to detect four infectious viral genome copies of MERS-CoV within a 60 min reaction time period. Significantly, EvaGreen dye has better signal read-out properties in one-pot RT-LAMP reaction and is more compatible with DNA polymerase than SYBR green I. Isothermally amplified specific N genes were further evaluated using field-deployable microchamber devices, leading to the specific identification of as few as 0.4 infectious viral genome copies, with no cross-reaction to the other acute respiratory disease viruses, including influenza type A (H1N1 and H3N2), type B, human coronavirus 229E, and human metapneumovirus. This sensitive, specific and feasible method provides a large-scale technical support in emergencies, and is also applied as a sample-to-detection module in Point of Care Testing devices.</p></abstract><kwd-group><kwd>one-pot reverse transcription (RT)</kwd><kwd>MERS-CoV</kwd><kwd>loop-mediated isothermal amplification (LAMP)</kwd><kwd>nucleocapsid (N) protein</kwd><kwd>EvaGreen</kwd></kwd-group><counts><fig-count count="8"></fig-count><table-count count="1"></table-count><equation-count count="0"></equation-count><ref-count count="31"></ref-count><page-count count="13"></page-count><word-count count="0"></word-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Pmc/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000C78 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd -nk 000C78 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Pmc
|étape= Curation
|type= RBID
|clé= PMC:5220095
|texte= One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Curation/RBID.i -Sk "pubmed:28119682" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |